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周期性介孔有机硅作为一种多功能纳米器件,用于大规模表征膜蛋白。

Periodic mesoporous organosilica as a multifunctional nanodevice for large-scale characterization of membrane proteins.

机构信息

Department of Chemistry, Institute of Biomedical Sciences, Fudan University, Shanghai 200433, China.

出版信息

Anal Chem. 2012 Jul 3;84(13):5809-15. doi: 10.1021/ac301146a. Epub 2012 Jun 12.

Abstract

A versatile protocol has been developed for large-scale characterization of hydrophobic membrane proteins based on the periodic mesoporous organosilica (PMO) acting as both an extractor for hydrophobic substrate capture and a nanoreactor for efficient in situ digestion. With introduction of organic groups in the pore frameworks and the presence of hydrophilic silanol groups on the surface, PMO can be well-dispersed into not only an organic solution to concentrate the dissolved membrane proteins but also an aqueous solution containing enzymes for sequential rapid proteolysis in the nanopores. The unique amphiphilic property of PMO ensures a facile switch in different solutions to realize the processes of substrate dissolution, enrichment, and digestion effectively. Furthermore, this novel PMO-assisted protocol has been successfully applied for identification of complex membrane proteins extracted from mouse liver as proof of general applicability.

摘要

一种基于周期性介孔有机硅(PMO)的多功能方案已被开发出来,用于大规模表征疏水性膜蛋白。PMO 既可以作为疏水性底物捕获的提取剂,又可以作为原位高效消化的纳米反应器。通过在孔骨架中引入有机基团和表面存在的亲水性硅醇基团,PMO 不仅可以很好地分散在有机溶剂中以浓缩溶解的膜蛋白,还可以分散在含有酶的水溶液中,以在纳米孔中进行连续的快速蛋白水解。PMO 的独特两亲性确保了在不同溶液之间的轻松切换,从而有效地实现了底物溶解、富集和消化的过程。此外,该新型 PMO 辅助方案已成功应用于从鼠肝中提取的复杂膜蛋白的鉴定,证明了其广泛适用性。

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